Journal Article

The E3 ligase HOIL-1 catalyses ester bond formation between ubiquitin and components of the Myddosome in mammalian cells

Knebel, Axel, Zhang, Jiazhen, Cohen, Philip, Kelsall, Ian R., Arthur, J. Simon C.

2019

Overview

The linear ubiquitin assembly complex (LUBAC) comprises 3 components: HOIP, HOIL-1, and Sharpin, of which HOIP and HOIL-1 are both members of the RBR subfamily of E3 ubiquitin ligases. HOIP catalyses the formation of Met1-linked ubiquitin oligomers (also called linear ubiquitin), but the function of the E3 ligase activity of HOIL-1 is unknown. Here, we report that HOIL-1 is an atypical E3 ligase that forms oxyester bonds between the C terminus of ubiquitin and serine and threonine residues in its substrates. Exploiting the sensitivity of HOIL-1–generated oxyester bonds to cleavage by hydroxylamine, and macrophages from knock-in mice expressing the E3 ligase-inactive HOIL-1[C458S] mutant, we identify IRAK1, IRAK2, and MyD88 as physiological substrates of the HOIL-1 E3 ligase during Toll-like receptor signaling. HOIL-1 is a monoubiquitylating E3 ubiquitin ligase that initiates the de novo synthesis of polyubiquitin chains that are attached to these proteins in macrophages. HOIL-1 also catalyses its own monoubiquitylation in cells and most probably the monoubiquitylation of Sharpin, in which ubiquitin is also attached by an oxyester bond. Our study establishes that oxyester-linked ubiquitylation is used as an intracellular signaling mechanism.

• Publisher: National Academy of Sciences
• Subject: Animals / Biochemistry / Biological Sciences / Catalysis / Gene Knock-In Techniques / Hydroxylamine / Immune system / Immunity, Innate / Interleukin-1 Receptor-Associated Kinases - metabolism / Intracellular signalling / IRAK protein / Macrophages / Macrophages - metabolism / Mammalian cells / Mice / Mice, Transgenic / Multiprotein Complexes - metabolism / Multiprotein Complexes - physiology / MyD88 protein / Oligomers / Proteins / Serine / Substrates / Threonine / Toll-like receptors / Ubiquitin / Ubiquitin - metabolism / Ubiquitin-protein ligase / Ubiquitin-Protein Ligases - metabolism / Ubiquitination